title: A new device for high-pressure freezing of cultured cell monolayer using 10-microm-thin stainless discs as both culture plate and specimen carrier.
abstract: High-pressure freezing (HPF) has been generally accepted as the most reliable method for cryofixation of biological samples, yielding a deep vitreous freezing. In recent cell biology, mammalian cultured cells are widely used, but HPF of cultured cell monolayer has not reached its full potential. In this study, we developed a new reliable device for HPF of cultured cell monolayer by using a 10-microm-thin stainless disc both as culture plate and specimen carrier. We describe the practical procedure, and demonstrate fine structures of HeLa cells cultured and cryofixed on the stainless discs as results.
authors.affiliation: Department of Anatomy, Ultrastructural Cell Biology, Faculty of Medicine, University of Miyazaki, Japan. sawa@fc.miyazaki-u.ac.jp
authors.forename: Akira
authors.initials: A
authors.lastname: Sawaguchi
authors.forename: Fumiyo
authors.initials: F
authors.lastname: Aoyama
authors.forename: Soyuki
authors.initials: S
authors.lastname: Ide
authors.forename: Yoshiteru
authors.initials: Y
authors.lastname: Goto
authors.forename: Tatsuo
authors.initials: T
authors.lastname: Suganuma
mesh_terms: D002478:Cells, Cultured / Q000648:ultrastructure*; D015925:Cryopreservation / Q000295:instrumentation / Q000379:methods; D004867:Equipment Design; D017110:Freeze Substitution; D006367:HeLa Cells; D006801:Humans; D008854:Microscopy, Electron; D011312:Pressure